PROJECT I. Sweat Patch Biomarkers: Clinical Studies. This is currently the main focus of the SNIB. We previously showed that neural and immune biomarkers are detectable in sweat and strongly correlate with plasma levels in a group of women with major depressive disorder (MDD) in clinical remission (Marques-Deak, 2006, J Immunol Methods; Cizza 2008, Biol Psych). Specifically, pro-inflammatory cytokines were elevated, as was the sympathetic neuropeptide neuropeptide Y (NPY) and the sensory/pain-related neuropeptides, substance P (SP) and CGRP (calcitonin gene-related peptide), while the parasympathetic neuropeptide vasoactive intestinal polypeptide (VIP) was significantly decreased. This pattern is consistent with a shift in MDD from parasympathetic to sympathetic tone, and an underlying pro-inflammatory state that could account for enhanced susceptibility to conditions known to be co-morbidly expressed with MDD, including cardiovascular disease, osteoporosis and diabetes. Moreover, biomarker levels strongly correlated with symptoms of depression and anxiety, indicating functional significance of these biomarker profiles. In order to determine the extent to which particular biomarker profiles reflect specific diseases or a state of health, in FY11 we are continuing to apply sweat patches in five ongoing IRB approved clinical protocols in collaboration with other NIH institutes and extramural institutions, including: (i) Emory University TRD-Infliximab Study. (Emory University IRB 00011734, NIMH OHSR Exemption #4025); (ii) Brazil - OCD Study (NIH protocol IRB 3737); (iii) Emory/CDC CFS Study (Emory University IRB 000551-2005, NIMH OHSR Exemption #4026); (iv) NCCAM - Tai Chi/Cancer Survivor Study (NCI protocol #06-AT-0016); (v) GSA - Work Environment Study (NIA IRB 2003-142). Results of study (v), (GSA Workplace Study) indicate that physical features of the workplace environment are associated with altered measures of the physiological stress response, as indicated by a greater rise in salivary cortisol upon awakening and flatter circadian variation of heart rate variability in subjects occupying old office space compared to those in new office space (Thayer et al. 2010 Eur J Cardiovasc Prev Rehabil). Our current studies focus on transitioning to a more high-throughput static antibody glass chip microarray platform and developing a better patch using nanotechnology. In FY10-11 the SNIB obtained funding support for this project through an NIH Directors Challenge Award in partnership with the NCCAM Director; NCCAM; the Center for Neuroscience and Regenerative Medicine (CNRM)/USUHS/DoD/NIH); and NIMH IRP funds. Project II Summary: Animal Models of Glucocorticoid Resistance, Inflammation and Behavior: In our studies evaluating the effects of the steroid hormone progesterone on immune cell (dendritic cell, DC) function, activation, and the role in inflammation and host defense, we assessed DCs from lymphoid (spleen, bone marrow, thymus) and non-lymphoid (liver, kidney, uterus) tissues. Our results show that progesterone in non-pregnancy-associated concentrations and through a progesterone receptor (PR)-mediated mechanism suppresses mature DC production of pro-inflammatory and helper T cell-related cytokines, cell surface marker expression (co-stimulatory molecules and chemokine receptors), cellular migration/motility, and stimulation of T cell proliferation but has little effect on immature DC antigen uptake (Butts et al. 2009 Methods Mol Biol; Butts et al. 2010 Mucosal Immunol). These effects, which are comparable to those of glucocorticoids on DC function, indicate that progesterone plays an important role in regulation of innate and adaptive immunity in females. We also found that progesterone effects on DC function vary throughout the rodent estrous cycle and are dependent on PR expression, which varies throughout the cycle in vitro and in vivo with intravaginal and intraperitoneal administration. This indicates that females' immune responses vary physiologically throughout the estrous cycle and has important implications for clinical susceptibility to infection and inflammation throughout the cycle and during pregnancy. Such physiological fluctuations in hormone status could also impact cellular responses that play a role in mood. Since cytokines are known to affect mood, and may play a role in some forms of depression as well as in mood alterations in sickness behavior, factors such as progesterone, which alter cytokine production by cells could contribute to differential mood disorder susceptibilities in females throughout the life cycle. Furthermore, progesterone has been shown to have beneficial therapeutic effects in traumatic brain injury (TBI) and post-traumatic stress disorder (PTSD). As such this data is important in understanding the pathogenesis and specific clinical conditions in which progesterone may be beneficial. In FY2011 we also showed tissue specific differences in glucocorticoid receptor (GR) expression in immune cells, which correlated with presence of tissue damage in a mouse model of West Nile virus infection (Butts et al. 2011 Brain Beh Immun.) This suggests that GR expression may play a role in severity of local inflammation in the brain and its pathological sequelae. In our animal studies in GRdim glucocorticoid receptor (GR) impaired mice we have found that an intact GR is essential for full recovery from inflammatory challenge. These animal studies are currently being concluded to allow SNIB to fully focus on Project 1: clinical translational sweat patch studies.